Department of Emergency Medicine, China-Japan Union Hospital of Jilin University, Changchun, Jilin, China.
Department of Critical Medicine, China-Japan Union Hospital of Jilin University, Changchun, Jilin, China.
Shock. 2024 Apr 1;61(4):620-629. doi: 10.1097/SHK.0000000000002245. Epub 2023 Nov 15.
Background: Sepsis is a systemic inflammatory disease that can cause multiple organ damage. Circular RNAs (circRNAs) have been reported to play a regulatory role in sepsis-induced acute kidney injury (AKI); however, the role of circ_0114428 has not been studied. Methods: In this study, HK2 cells were treated with different concentrations of LPS to induce cell damage, and then the expressions of circ_0114428, microRNA-215-5p (miR-215-5p), and tumor necrosis factor receptor-associated factor 6 (TRAF6) were detected by quantitative real-time polymerase chain reaction (qRT-PCR), and Western blot examined the Bax and cleaved-Caspase-3 proteins. Cell proliferation was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and thymidine analog 5-ethynyl-2'-deoxyuridine (EdU) assay. In addition, cell apoptosis was detected by flow cytometry, and the levels of inflammatory factors were detected by enzyme-linked immunosorbent assay. Results: After LPS treatment with different concentrations, we found that LPS at 10 μg/mL had the best effect on HK2 cells. Circ_0114428 was highly expressed in sepsis-AKI patients and LPS-treated HK2 cells. Knockdown of circ_0114428 restored the effects of LPS treatment on proliferation, apoptosis, and inflammatory response of HK2 cells. MiR-215-5p was a target of circ_0114428, and TRAF6 was a downstream target of miR-215-5p. Circ_0114428 regulated TRAF6 expression by sponging miR-215-5p in LPS-treated HK2 cells. Circ_0114428 regulated LPS-induced NF-κB signaling in HK2 cells by targeting miR-215-5p/TRAF6 axis. Conclusion: Circ_0114428 knockdown abolished the cell proliferation, apoptosis, and inflammatory damage in LPS-induced HK2 cells by targeting miR-215-5p/TRAF6/NF-κB.
败血症是一种全身性炎症性疾病，可导致多器官损伤。环状 RNA（circRNA）已被报道在败血症诱导的急性肾损伤（AKI）中发挥调节作用；然而，circ_0114428 的作用尚未研究。方法：在这项研究中，用不同浓度的 LPS 处理 HK2 细胞以诱导细胞损伤，然后通过定量实时聚合酶链反应（qRT-PCR）检测 circ_0114428、microRNA-215-5p（miR-215-5p）和肿瘤坏死因子受体相关因子 6（TRAF6）的表达，并用 Western blot 检测 Bax 和 cleaved-Caspase-3 蛋白。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴盐（MTT）试验和胸苷类似物 5-乙炔基-2'-脱氧尿苷（EdU）试验检测细胞增殖。此外，通过流式细胞术检测细胞凋亡，通过酶联免疫吸附试验检测炎症因子水平。结果：用不同浓度的 LPS 处理后，我们发现 10 μg/mL 的 LPS 对 HK2 细胞的效果最佳。circ_0114428 在败血症-AKI 患者和 LPS 处理的 HK2 细胞中高表达。circ_0114428 的敲低恢复了 LPS 处理对 HK2 细胞增殖、凋亡和炎症反应的影响。miR-215-5p 是 circ_0114428 的靶标，TRAF6 是 miR-215-5p 的下游靶标。circ_0114428 通过海绵吸附 LPS 处理的 HK2 细胞中的 miR-215-5p 来调节 TRAF6 的表达。circ_0114428 通过靶向 miR-215-5p/TRAF6 轴调节 LPS 诱导的 HK2 细胞中的 NF-κB 信号通路。结论：circ_0114428 的敲低通过靶向 miR-215-5p/TRAF6/NF-κB 消除了 LPS 诱导的 HK2 细胞中的细胞增殖、凋亡和炎症损伤。
